Research on methods of analytical ultracentrifugation is proposed. The areas of endeavor include: (1) Development of methods to determine point-by-point molecular weight averages in sedimentation velocity experiments. (2) Use of difference sedimentation velocity experiments to determine self-association and complex formation in impure protein mixtures. (3) Use of difference sedimentation equilibrium methods to determine stoichiometry and, perhaps, equilibrium constants of self-associating systems in impure mixtures. (4) Further study of the active enzyme sedimentation technique to determine molecular weights of active species and detection of minor active components. This methodology as well as established techniques are to be used to study the self-association of alpha-chymotrypsin at high pH, the mechanism of colchicine disruption of polymerized tubulin, and the self-associating IgG rheumatoid factors of rheumatoid arthritis. In addition to these projects it is proposed to digest dissociated enzymes with exopeptidases in order to create small, enzymatically active protein species. We propose to continue studies of the evolution of proteins of diverse structure and function. Computer methods are used to align the primary sequences of families of proteins. Work on the statistical evaluation of the alignment is proposed. Extension to assessment of 3 dimensional similarity is proposed. Studies of translational frictional coefficients of macromolecules are in progress. Use of a test sphere shell of 1.4 A test spheres together with the Kirkwood pearl-necklace formula allows computation of the frictional coefficient of known proteins. More exact computations using recent theories is proposed. BIBLIOGRAPHIC REFERENCES: Teller, D.C. and Deranleau, D.A.: Direct Determination of the Stoichiometry of charge transfer Complexes. Biochem. Biophys. Acta 421, 416-419 (1976). Kurachi, K., Schmer, G., Hermodson, M.A., Teller, D.C. and Davie, E.W.: Characterization of Human, Bovine, and Horse Antithrombin III. Biochemistry 15, 368-373 (1976).